JP2015514404A5 - - Google Patents
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- JP2015514404A5 JP2015514404A5 JP2015506048A JP2015506048A JP2015514404A5 JP 2015514404 A5 JP2015514404 A5 JP 2015514404A5 JP 2015506048 A JP2015506048 A JP 2015506048A JP 2015506048 A JP2015506048 A JP 2015506048A JP 2015514404 A5 JP2015514404 A5 JP 2015514404A5
- Authority
- JP
- Japan
- Prior art keywords
- polynucleotide
- birds
- embryo
- germ cells
- transfection reagent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 241000271566 Aves Species 0.000 claims 15
- 229920000023 polynucleotide Polymers 0.000 claims 12
- 239000002157 polynucleotide Substances 0.000 claims 12
- 210000001161 Embryo, Mammalian Anatomy 0.000 claims 10
- 210000004602 germ cell Anatomy 0.000 claims 9
- 239000012096 transfection reagent Substances 0.000 claims 7
- 239000000203 mixture Substances 0.000 claims 5
- 238000001890 transfection Methods 0.000 claims 4
- 229920000160 (ribonucleotides)n+m Polymers 0.000 claims 3
- 210000004204 Blood Vessels Anatomy 0.000 claims 3
- PFNFFQXMRSDOHW-UHFFFAOYSA-N Spermine Natural products NCCCNCCCCNCCCN PFNFFQXMRSDOHW-UHFFFAOYSA-N 0.000 claims 3
- 230000023298 conjugation with cellular fusion Effects 0.000 claims 3
- 230000013011 mating Effects 0.000 claims 3
- 230000000051 modifying Effects 0.000 claims 3
- 230000021037 unidirectional conjugation Effects 0.000 claims 3
- 125000002091 cationic group Chemical group 0.000 claims 2
- -1 cationic lipid Chemical class 0.000 claims 2
- 210000004027 cells Anatomy 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 229920001184 polypeptide Polymers 0.000 claims 2
- 229940063675 spermine Drugs 0.000 claims 2
- 230000029812 viral genome replication Effects 0.000 claims 2
- LDGWQMRUWMSZIU-UHFFFAOYSA-M 2,3-bis(octadec-9-enoxy)propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCCC=CCCCCCCCCOCC(C[N+](C)(C)C)OCCCCCCCCC=CCCCCCCCC LDGWQMRUWMSZIU-UHFFFAOYSA-M 0.000 claims 1
- LDGWQMRUWMSZIU-LQDDAWAPSA-M 2,3-bis[(Z)-octadec-9-enoxy]propyl-trimethylazanium;chloride Chemical group [Cl-].CCCCCCCC\C=C/CCCCCCCCOCC(C[N+](C)(C)C)OCCCCCCCC\C=C/CCCCCCCC LDGWQMRUWMSZIU-LQDDAWAPSA-M 0.000 claims 1
- KWVJHCQQUFDPLU-YEUCEMRASA-N 2,3-bis[[(Z)-octadec-9-enoyl]oxy]propyl-trimethylazanium Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(C[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC KWVJHCQQUFDPLU-YEUCEMRASA-N 0.000 claims 1
- WALUVDCNGPQPOD-UHFFFAOYSA-M 2,3-di(tetradecoxy)propyl-(2-hydroxyethyl)-dimethylazanium;bromide Chemical compound [Br-].CCCCCCCCCCCCCCOCC(C[N+](C)(C)CCO)OCCCCCCCCCCCCCC WALUVDCNGPQPOD-UHFFFAOYSA-M 0.000 claims 1
- 241000272525 Anas platyrhynchos Species 0.000 claims 1
- 241000272814 Anser sp. Species 0.000 claims 1
- RBGCWNFXPQALRC-UHFFFAOYSA-N CN(CCCN(CCCCN(CCC(N(C(CCCCCCCCCCCCCCC)=O)C(CCCCCCCCCCCCCCC)=O)(C(CCCCCCCCCCCCCCC)=O)C(CCCCCCCCCCCCCCC)=O)C)C)C Chemical compound CN(CCCN(CCCCN(CCC(N(C(CCCCCCCCCCCCCCC)=O)C(CCCCCCCCCCCCCCC)=O)(C(CCCCCCCCCCCCCCC)=O)C(CCCCCCCCCCCCCCC)=O)C)C)C RBGCWNFXPQALRC-UHFFFAOYSA-N 0.000 claims 1
- QPYUGOUSOOCGGQ-UHFFFAOYSA-N CN(CCCN(CCCCN(CCC(N(CCCCCCCCCCCC)CCCCCCCCCCCC)(CCCCCCCCCCCC)CCCCCCCCCCCC)C)C)C Chemical compound CN(CCCN(CCCCN(CCC(N(CCCCCCCCCCCC)CCCCCCCCCCCC)(CCCCCCCCCCCC)CCCCCCCCCCCC)C)C)C QPYUGOUSOOCGGQ-UHFFFAOYSA-N 0.000 claims 1
- NFIDGYIHSCPMMK-SDLHUVDUSA-N CN(CCCN(CCCCN(CCC(N(CCCCCCCC\C=C/CCCCCCCC)CCCCCCCC\C=C/CCCCCCCC)(CCCCCCCC\C=C/CCCCCCCC)CCCCCCCC\C=C/CCCCCCCC)C)C)C Chemical compound CN(CCCN(CCCCN(CCC(N(CCCCCCCC\C=C/CCCCCCCC)CCCCCCCC\C=C/CCCCCCCC)(CCCCCCCC\C=C/CCCCCCCC)CCCCCCCC\C=C/CCCCCCCC)C)C)C NFIDGYIHSCPMMK-SDLHUVDUSA-N 0.000 claims 1
- 229920000033 CRISPR Polymers 0.000 claims 1
- 238000010354 CRISPR gene editing Methods 0.000 claims 1
- XULFJDKZVHTRLG-JDVCJPALSA-N DOSPA trifluoroacetate Chemical group [O-]C(=O)C(F)(F)F.CCCCCCCC\C=C/CCCCCCCCOCC(C[N+](C)(C)CCNC(=O)C(CCCNCCCN)NCCCN)OCCCCCCCC\C=C/CCCCCCCC XULFJDKZVHTRLG-JDVCJPALSA-N 0.000 claims 1
- 210000003278 Egg Shell Anatomy 0.000 claims 1
- 241000287828 Gallus gallus Species 0.000 claims 1
- 101700080605 NUC1 Proteins 0.000 claims 1
- 229920001850 Nucleic acid sequence Polymers 0.000 claims 1
- 241000286209 Phasianidae Species 0.000 claims 1
- 108020004459 Small Interfering RNA Proteins 0.000 claims 1
- 229920001891 Small hairpin RNA Polymers 0.000 claims 1
- 238000010459 TALEN Methods 0.000 claims 1
- DTQVDTLACAAQTR-UHFFFAOYSA-M Trifluoroacetate Chemical compound [O-]C(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-M 0.000 claims 1
- 241000700605 Viruses Species 0.000 claims 1
- 230000000840 anti-viral Effects 0.000 claims 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 claims 1
- UMGXUWVIJIQANV-UHFFFAOYSA-M didecyl(dimethyl)azanium;bromide Chemical compound [Br-].CCCCCCCCCC[N+](C)(C)CCCCCCCCCC UMGXUWVIJIQANV-UHFFFAOYSA-M 0.000 claims 1
- PSLWZOIUBRXAQW-UHFFFAOYSA-M dimethyl(dioctadecyl)azanium;bromide Chemical compound [Br-].CCCCCCCCCCCCCCCCCC[N+](C)(C)CCCCCCCCCCCCCCCCCC PSLWZOIUBRXAQW-UHFFFAOYSA-M 0.000 claims 1
- 235000013601 eggs Nutrition 0.000 claims 1
- 210000002257 embryonic structures Anatomy 0.000 claims 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims 1
- 238000002347 injection Methods 0.000 claims 1
- 239000007924 injection Substances 0.000 claims 1
- 150000002632 lipids Chemical class 0.000 claims 1
- 230000001264 neutralization Effects 0.000 claims 1
- 101700006494 nucA Proteins 0.000 claims 1
- 125000002811 oleoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])/C([H])=C([H])\C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims 1
- 125000001820 oxy group Chemical group [*:1]O[*:2] 0.000 claims 1
- 239000002924 silencing RNA Substances 0.000 claims 1
- 239000004055 small Interfering RNA Substances 0.000 claims 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims 1
- 239000011701 zinc Substances 0.000 claims 1
- 229910052725 zinc Inorganic materials 0.000 claims 1
Claims (16)
(i)トランスフェクション試薬と混合したポリヌクレオチドを含むトランスフェクション混合物を鳥類胚の血管に注入する工程
を含み、それによってポリヌクレオチドが鳥類における1個または複数の生殖細胞のゲノムに挿入される方法。 A method for producing a bird containing genetically modified germ cells, comprising:
(i) A method comprising injecting a transfection mixture comprising a polynucleotide mixed with a transfection reagent into a blood vessel of an avian embryo, whereby the polynucleotide is inserted into the genome of one or more germ cells in the bird.
をさらに含む、請求項1に記載の方法。 2. The method of claim 1, further comprising (ii) incubating the embryo at a temperature sufficient for the embryo to develop in the chick.
(ii) トランスフェクション試薬がDOSPA(2,3-ジオレイルオキシ-N-[2(スペルミンカルボキサミド)エチル]-N,N-ジメチル-1-プロパンアミニウムトリフルオロアセテート)、 DOSPER(1,3-ジオレオイルオキシ-2-(6カルボキシスペルミル)-プロピルアミド)、TMTPS(テトラメチルテトラパルミトイルスペルミン)、TMTOS(テトラメチルテトラオレイルスペルミン)、TMTLS(テトラメチルテトララウリルスペルミン)、TMTMS(テトラメチルテトラミリスチルスペルミン)およびTMDOS(テトラメチルジオレイルスペルミン)から選択される1つまたは複数の多価のカチオン性脂質を含む、請求項4に記載の方法。 (i) The transfection reagent is DOTMA (N- [1- (2,3-dioleoyloxy) -propyl] -N, N, N-trimethylammonium chloride), DOTAP (1,2-bis (oleoyl One selected from (oxy) -3-3- (trimethylammonium) propane), DMRIE (1,2-dimyristyloxypropyl-3-dimethyl-hydroxyethylammonium bromide) and DDAB (dimethyldioctadecylammonium bromide) or including the cationic lipids of the plurality of monovalent and / or
(ii) The transfection reagent is DOSPA (2,3-dioleyloxy-N- [2 (sperminecarboxamido) ethyl] -N, N-dimethyl-1-propaneaminium trifluoroacetate), DOSPER (1,3- (Dioleoyloxy-2- (6carboxyspermyl) -propylamide), TMPTS (tetramethyltetrapalmitoylspermine), TMTOS (tetramethyltetraoleylspermine), TMTLS (tetramethyltetralaurylspermine), TTMMS (tetramethyltetramine) 5. The method of claim 4, comprising one or more polyvalent cationic lipids selected from myristyl spermine) and TMDOS (tetramethyldiolyl spermine) .
(i)トランスフェクション試薬と混合したポリヌクレオチドを含むトランスフェクション混合物を卵内に含有される鳥類胚の血管に注入する工程、および
(ii)胚の雛への発生を可能にするために十分な温度で胚をインキュベートする工程
を含み、
ポリヌクレオチドが鳥類における1個または複数の生殖細胞のゲノムに挿入される方法。 A method for genetically modifying germ cells in birds,
(i) injecting a transfection mixture comprising a polynucleotide mixed with a transfection reagent into a blood vessel of an avian embryo contained in an egg; and
(ii) incubating the embryo at a temperature sufficient to allow development of the embryo into chicks,
A method wherein a polynucleotide is inserted into the genome of one or more germ cells in birds.
(i)請求項1から10のいずれか一項に記載の方法を用いて作製された、遺伝子改変生殖細胞を含む鳥類を得る工程、
(ii)遺伝子改変生殖細胞を含む鳥類から交配により子孫を作製する工程、および
(iii)ゲノムに挿入されたポリヌクレオチドを含む子孫を選択する工程
を含む方法。 A method for producing genetically modified birds,
(i) obtaining a bird comprising a genetically modified germ cell, produced using the method according to any one of claims 1 to 10 ,
(ii) step of producing progeny by mating from birds comprising genetically modified germ cells, and
(iii) A method comprising the step of selecting progeny containing a polynucleotide inserted into the genome.
(i)請求項1から12のいずれか一項に記載の方法を実施して、雛または子孫を作製する工程
(ii)雛または子孫を性的に成熟した鳥類へと育てる工程、および
(iii)性的に成熟した鳥類から交配により遺伝子改変鳥類を作製する工程
を含む方法。 A method of mating genetically modified birds,
(i) carrying out the method according to any one of claims 1 to 12 to produce chicks or offspring
(ii) raising chicks or offspring into sexually mature birds; and
which comprises the step of producing a genetically modified birds by mating from (iii) sexually mature birds.
(i)トランスフェクション試薬と混合したポリヌクレオチドを含むトランスフェクション混合物を鳥類胚の血管に注入し、それによってポリヌクレオチドが鳥類における1個または複数の生殖細胞のゲノムに挿入される工程、および
(ii)胚の雛への発生を可能にするために十分な温度で胚をインキュベートする工程
を含み、ポリヌクレオチドが鳥類における形質をモジュレートするポリペプチドまたは二本鎖領域を含むRNA分子をコードする方法。 A method for modulating traits in birds, comprising:
(i) injecting a transfection mixture comprising a polynucleotide mixed with a transfection reagent into a blood vessel of an avian embryo, whereby the polynucleotide is inserted into the genome of one or more germ cells in the bird; and
(ii) incubating the embryo at a temperature sufficient to allow development into embryonic chicks, wherein the polynucleotide encodes an RNA molecule comprising a polypeptide or double-stranded region that modulates a trait in birds how to.
Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201261636331P | 2012-04-20 | 2012-04-20 | |
| US61/636,331 | 2012-04-20 | ||
| US201361783823P | 2013-03-14 | 2013-03-14 | |
| US61/783,823 | 2013-03-14 | ||
| AU2013204327 | 2013-04-12 | ||
| AU2013204327A AU2013204327B2 (en) | 2012-04-20 | 2013-04-12 | Cell transfection method |
| PCT/AU2013/000414 WO2013155572A1 (en) | 2012-04-20 | 2013-04-19 | Cell transfection method |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2015514404A JP2015514404A (en) | 2015-05-21 |
| JP2015514404A5 true JP2015514404A5 (en) | 2016-06-16 |
| JP6531239B2 JP6531239B2 (en) | 2019-06-19 |
Family
ID=49515073
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2015506048A Active JP6531239B2 (en) | 2012-04-20 | 2013-04-19 | Cell transfection method |
Country Status (9)
| Country | Link |
|---|---|
| US (4) | US10897881B2 (en) |
| EP (2) | EP3460065B1 (en) |
| JP (1) | JP6531239B2 (en) |
| KR (1) | KR102076784B1 (en) |
| CN (2) | CN104619848B (en) |
| AU (2) | AU2013204327B2 (en) |
| BR (1) | BR112014026186B8 (en) |
| ES (2) | ES2704155T3 (en) |
| WO (1) | WO2013155572A1 (en) |
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